Enhanced gametocyte formation in young erythrocytes by Plasmodium falciparum in vitro.

Two gametocyte-forming clones, HB-3 and 3D7, were used. Concentrates of late stage parasites were mixed with bloods containing different proportions of young erythrocytes, and the parasitemia and proportion of gametocytes determined after 2, 3 or 4 days of culture. Significantly more gametocytes were formed in light cells than in heavy cells separated from the same normal blood samples. Up to seven times more gametocytes were formed in reticulocyte-rich bloods from patients with sickle cell anemia than in normal control blood.     

Levels of metallic mercury and mercuric ion in the venous and arterial bloods of normal and acatalasemic mice following exposure to mercury vapor

Levels of metallic mercury and mercuric ion in the arterial and venous bloods of normal and acatalasemic mice exposed to metallic mercury vapor in vitro and in vivo were investigated. Mercury uptake in venous blood from air saturated with mercury vapor with or without hydrogen peroxide in vitro was determined. Level of mercuric ion in venous blood of normal mice was significantly higher than that of acatalasemic mice. By contrast, metallic mercury in venous blood of acatalasemic mice was elevated relative to level in normal mice. Metallic mercury level in red blood cells and plasma was also significantly higher in acatalasemic mice. The ratio of metallic mercury to total mercury (Hg degrees + Hg2+) in the arterial and venous bloods of acatalasemic mice exposed to metallic mercury vapor was increased relative to normal mice. This ratio in red blood cells and plasma in the venous bloods of acatalasemic mice in vivo was also significantly higher than those of normal mice. The significance of metallic mercury in plasma for distribution of mercury in organs is discussed.     

Differentiation of Gametocytes in Microcultures of Human Blood Infected with Plasmodium falciparum*

SYNOPSIS. Gametocytes differentiated from ring-stage parasites in microcultures of human blood infected with Plasmodium falciparum. Immature gametocytes could be distinguished morphologically from late asexual trophozoites after ～ 40 h of culture. Differentiation into crescentic forms took several days and the incorporation of [³H]-isoleucine by developing gametocytes was demonstrated. About 1% of red cells contained gametocytes at the maximum densities attained. Differentiation of gametocytes occurred either directly from rings placed in culture or from the progeny of subsequent cycles of schizogony and invasion in vitro. The latter occurrence was confirmed by the development of gametocytes in marker fetal red cells added to cultures, although fetal red cells provide a less favorable environment than those with HbA for growth of the parasites.     

A New Saurian Malarial Parasite Plasmodium balli from Panama

SYNOPSIS. Plasmodium balli sp. nov. is described from Anolis lionotus and A. poecilopus of central Panama.
Large, elongate gametocytes and segmenters containing up to 100 merozoites are produced by P. balli. Proerythrocytes and normoblasts are more commonly parasitized than erythrocytes. Pigment is uncommon, but when present consists of a minute dot. Hypertrophy, distortion and lysis of host cell nuclei may result from parasitization of immature blood cells by gametocytes, while enucleated host cells are common.     

Elongate and Round Gametocytes of Leucocytozoon simondi (Mathis & Léger) in Ducks Inoculated with Megaloschizonts

SYNOPSIS. Single megaloschizonts give rise to elongate and round gametocytes, the former outnumbering the latter. Male and female elongate gametocytes develop from merozoites of a single megaloschizont. Elongate gametocytes were seen 2–7 days and round gametocytes 6–11 days after megaloschizonts had been inoculated into ducklings. Experimental evidence indicates that merozoites of megaloschizonts invade blood cells and develop into elongate gametocytes. Other merozoites infect tissue cells and develop into secondary exoerythrocytic schizonts which give rise to round gametocytes. Relapse in Leucocytozoon simondi infections is discussed in relation to megaloschizont-induced exoerythrocytic schizogony.     

Schizogony and Gametogony of Leucocytozoon simondi and Associated Reactions in the Avian Host*

SYNOPSIS. Stages of development of Leucocytozoon simondi in White Pekin ducklings and their reactions to the parasite were studied on successive days after infecting them artificially with sporozoites from Simulium rugglesi. The minimum prepatent period was 5 days. The first asexual cycle occurred exclusively in the parenchymal cells of the liver. Progeny of these hepatic schizonts followed one of 3 courses: (a) invaded parenchymal liver cells to give rise to another hepatic cycle, (b) penetrated blood cells to form round gametocytes, and (c) were phagocytized by macrophages and grew into megaloschizonts thruout the body. The appearance of elongating gametocytes coincided with the period of maturation and release of merozoites from the megaloschizonts. Experimental evidence supports the hypothesis that the round gametocytes arise from the hepatic schizonts and the elongate forms from the megaloschizonts. Mature megaloschizonts released millions of merozoites, but a high 2nd peak in parasitemia did not develop because of retention of developing gametocytes in the deep circulation, particularly the liver and spleen, and a pronounced host reaction.     

Human T clones reactive to the sexual stages of Plasmodium falciparum malaria. High frequency of gamete-reactive T cells in peripheral blood from nonexposed donors

Malarial gametocytes, which are taken up by mosquitoes during a blood meal, develop in the gut of the mosquito into gametes. Gametes and gametocytes contain the target antigens of transmission-blocking immunity. Here, we show that the peripheral blood of nonexposed donors contains Plasmodium falciparum gamete-reactive T cells at frequencies ranging from 1/300 to 1/4000. Studies on long-term clones demonstrated that these cells often recognized antigens shared between gametes and asexual stage parasites or even between heterologous gametes, although it has been possible to derive a P. falciparum gamete-specific T clone. The T clones examined were T3+, T4+, T8-, and either HLA-DR- or HLA-DQ-restricted. They responded to gametes by both proliferation and the secretion of gamma-interferon. The gamete-specific clone and other asexual cross-reactive clones examined could be stimulated in vitro by a preparation of mature gametocytes within RBC, but not by RBC alone, suggesting that gametocytes are immunogenic or can become immunogenic for T cells in vivo. The significance of these observations to mosquito transmission of malaria and development and application of a gamete vaccine are discussed.     

Host cell deformability is linked to transmission in the human malaria parasite Plasmodium falciparum

Gametocyte maturation in Plasmodium falciparum is a critical step in the transmission of malaria. While the majority of parasites proliferate asexually in red blood cells, a small fraction of parasites undergo sexual conversion and mature over 2 weeks to become competent for transmission to a mosquito vector. Immature gametocytes sequester in deep tissues while mature stages must be able to circulate, pass the spleen and present themselves to the mosquito vector in order to complete transmission. Sequestration of asexual red blood cell stage parasites has been investigated in great detail. These studies have demonstrated that induction of cytoadherence properties through specific receptor-ligand interactions coincides with a significant increase in host cell stiffness. In contrast, the adherence and biophysical properties of gametocyte-infected red blood cells have not been studied systematically. Utilizing a transgenic line for 3D live imaging, in vitro capillary assays and 3D finite element whole cell modelling, we studied the role of cellular deformability in determining the circulatory characteristics of gametocytes. Our analysis shows that the red blood cell deformability of immature gametocytes displays an overall decrease followed by rapid restoration in mature gametocytes. Intriguingly, simulations suggest that along with deformability variations, the morphological changes of the parasite may play an important role in tissue distribution in vivo. Taken together, we present a model, which suggests that mature but not immature gametocytes circulate in the peripheral blood for uptake in the mosquito blood meal and transmission to another human host thus ensuring long-term survival of the parasite.     

Differential adhesive properties of sequestered asexual and sexual stages of Plasmodium falciparum on human endothelial cells are tissue independent

The protozoan parasite Plasmodium falciparum, responsible for the most severe form of malaria, is able to sequester from peripheral circulation during infection. The asexual stage parasites sequester by binding to endothelial cell receptors in the microvasculature of various organs. P. falciparum gametocytes, the developmental stages responsible for parasite transmission from humans to Anopheles mosquitoes, also spend the almost ten days necessary for their maturation sequestered away from the peripheral circulation before they are released in blood mainstream. In contrast to those of asexual parasites, the mechanisms and cellular interactions responsible for immature gametocyte sequestration are largely unexplored, and controversial evidence has been produced so far on this matter. Here we present a systematic comparison of cell binding properties of asexual stages and immature and mature gametocytes from the reference P. falciparum clone 3D7 and from a patient parasite isolate on a panel of human endothelial cells from different tissues. This analysis includes assays on human bone marrow derived endothelial cell lines (HBMEC), as this tissue has been proposed as a major site of gametocyte maturation. Our results clearly demonstrate that cell adhesion of asexual stage parasites is consistently more efficient than that, virtually undetectable of immature gametocytes, irrespectively of the endothelial cell lines used and of parasite genotypes. Importantly, immature gametocytes of both lines tested here do not show a higher binding efficiency compared to asexual stages on bone marrow derived endothelial cells, unlike previously reported in the only study on this issue. This indicates that gametocyte-host interactions in this tissue are unlikely to be mediated by the same adhesion processes to specific endothelial receptors as seen with asexual forms.     

Malarial Parasites of the "Jesu Cristo" Lizard Basiliscus basiliscus (Iguanidae) in Panama

SYNOPSIS. The iguanid lizard Basiliscus basiliscus in Panama is parasitized by Plasmodium basilisci and P. achiotense sp. nov. P. basilisci in this host is characterized by schizonts containing 4–14 merozoites, with schizonts parasitizing proerythrocytes containing more merozoites than those in erythrocytes. Asexual parasites lack cytoplasmic projections, while mature gametocytes are round or oval with regular margins.
P. achiotense is characterized by the combination of prominently pigmented, large schizonts containing 36–56 merozoites and oval or round gametocytes which are about 1/3 larger than those of P. basilisci.
EE-schizonts of P. basilisci were observed commonly in thrombocytes and occasionally in lymphocytes, and appeared early in experimental infections induced by blood inoculation.     

Plasmodium cynomolgi: serum-mediated blocking and enhancement of infectivity to mosquitoes during infections in the natural host, Macaca sinica

The infectivity of Plasmodium cynomolgi in its natural host, the toque monkey, Macaca sinica, to Anopheles tessellatus mosquitoes was studied in relation to the evolution of anti-sexual-stage immunity in the host during the course of a blood-induced infection. The effects of serum on the infectivity of gametocytes and the intrinsic infectivity of gametocytes to mosquitoes on each day were assessed in membrane feeding experiments. Mosquitoes were also directly fed on the animal on each day. Our results demonstrate that during the very early patent period, before the peak of gametocytemia, the infection serum enhanced the infectivity of gametocytes up to two to three times above their infectivity in normal monkey serum. Subsequently, serum drawn post-peak of parasitemia ceased to enhance, and began to suppress, infectivity. After 2-3 months, long after parasitemias ceased patency, the serum no longer suppressed and between 3 and 4 months the serum again tended to enhance gamete infectivity before losing any significant effect. Serum infectivity enhancing effects were consistent with low indirect immunofluorescence test antibody titers against blood stage parasites first during the very early days of a blood infection before reaching blocking levels, and again during convalescence when antibodies were declining. The serum infectivity blocking effects on gametocytes were seen at the peak of antibody titers from about Days 9 to 23 of an infection. From 78 to 95% of the total infectivity of the parasite to mosquitoes during an infection occurred when infectivity enhancing activity was present in the serum. Hence, the infectivity of the parasite to mosquitoes was largely dependent on infectivity enhancing antibodies in host serum.     

Studies of growth and development of gametocytes in Haemoproteus columbae Kruse.

As determined by their ability to exflagellate and round up, it took the macrogametocyte and the microgametocyte of Haemoproteus columbae in pigeons 68 and 116 h, respectively, after patency to reach maturity. Pigment granules appeared in the undifferentiated gametocytes 8 h after invasion of blood. Vacuoles were observed in young gametocytes and persisted in the older forms. The growth curve of H. columbae is close to the sigmoidal curve for growth in protozoa. Multiple infection was noticed in pigeons with high levels of parasitemia, but no more than 2 gametocytes reached maturity; such multiple infections were rare in relapses. The sex ratio of the gametocytes was 1:1. Strong lateral displacement of the nuclei of infected erythrocytes was the rule; hypertrophy was negligible.     

Studies of Growth and Development of Gametocytes in Haemoproteus columbae Kruse*

SYNOPSIS As determined by their ability to exflagellate and round up, it took the macrogametocyte and the microgametocyte of Haemoproteus columbae in pigeons 68 and 116 h, respectively, after patency to reach maturity. Pigment granules appeared in the undifferentiated gametocytes 8 h after invasion of blood. Vacuoles were observed in young gametocytes and persisted in the older forms. The growth curve of H. columbae is close to the sigmoidal curve for growth in protozoa. Multiple infection was noticed in pigeons with high levels of parasitemia, but no more than 2 gametocytes reached maturity; such multiple infections were rare in relapses. The sex ratio of the gametocytes was 1:1. Strong lateral displacement of the nuclei of infected erythrocytes was the rule; hypertrophy was negligible.     

Saurian Malarial Parasites in Eastern Panama

SYNOPSIS Plasmodium gonatodi sp. nov. is described from Gonatodes albogularis fuscus of eastern Panama. It is characterized by elongate gametocytes and polymorphic schizonts containing 12-46 nuclei when apparently mature. Both proerythrocytes and erythrocytes are commonly parasitized, host cells are hypertrophied and distorted, and their nuclei are displaced. Prematuration sexual stages may be irregularly shaped and larger than mature gametocytes.
Plasmodium diploglossi Aragão and Neiva , 1909 is reported from Mabuya mabouya in eastern Panama, and Plasmodium morulum sp. nov. is described from this host. P. morulum usually parasitizes immature erythrocytes, and is characterized by lenticular or oval to round gametocytes, and schizonts with 14-40 nuclei usually arranged in a globular mass. Host cells are slightly hypertrophied and distorted, and their nuclei are usually displaced. Inoculation of infected blood into clean hosts produces numerous schizonts in white cells as well as in the erythrocyte series.
Pigment in both P. gonatodi and P. morulum , if present, consists of a few minute dark dots which do not meet the polarized light test for hemozoin.     

Malaria-induced reduction of fecundity during the first gonotrophic cycle of Anopheles Stephensi mosquitoes

Abstract. Anopheles stephensi mosquitoes which had fed upon mice infected with Plasmodium yoelii nigeriensis malaria parasites produced significantly fewer eggs than mosquitoes fed on an uninfected mouse. Fecundity reduction was more pronounced when the bloodmeal contained malaria gametocytes and the mosquitoes developed oocysts. Egg production and haematin excretion were correlated for uninfected bloodfed mosquitoes; the presence of P.y. nigeriensis in the blood affected this relationship. Reduced fecundity was associated with a significant reduction of bloodmeal size (measured by haematin excretion) in mosquitoes which ingested gametocytaemic blood. The bloodmeal size in mosquitoes fed on parasitaemic blood without gametocytes was not significantly reduced. The use of haematin assays for determination of bloodmeal size in mosquitoes is discussed.     

Early gametocytes of the malaria parasite Plasmodium falciparum specifically remodel the adhesive properties of infected erythrocyte surface

In Plasmodium falciparum infections the parasite transmission stages, the gametocytes, mature in 10 days sequestered in internal organs. Recent studies suggest that cell mechanical properties rather than adhesive interactions play a role in sequestration during gametocyte maturation. It remains instead obscure how sequestration is established, and how the earliest sexual stages, morphologically similar to asexual trophozoites, modify the infected erythrocytes and their cytoadhesive properties at the onset of gametocytogenesis. Here, purified P. falciparum early gametocytes were used to ultrastructurally and biochemically analyse parasite‐induced modifications on the red blood cell surface and to measure their functional consequences on adhesion to human endothelial cells. This work revealed that stage I gametocytes are able to deform the infected erythrocytes like asexual parasites, but do not modify its surface with adhesive ‘knob’ structures and associated proteins. Reduced levels of the P. falciparum erythrocyte membrane protein 1 (PfEMP1) adhesins are exposed on the red blood cell surface bythese parasites, and the expression of the var gene family, which encodes 50–60 variants of PfEMP1, is dramatically downregulated in the transition from asexual development to gametocytogenesis. Cytoadhesion assays show that such gene expression changes and host cell surface modifications functionally result in the inability of stage I gametocytes to bind the host ligands used by the asexual parasite to bind endothelial cells. In conclusion, these results identify specific differences in molecular and cellular mechanisms of host cell remodelling and in adhesive properties, leading to clearly distinct host parasite interplays in the establishment of sequestration of stage I gametocytes and of asexual trophozoites.     

The novel putative transporter NPT1 plays a critical role in early stages of Plasmodium berghei sexual development

Transmission of Plasmodium species from a mammalian host to the mosquito vector requires the uptake, during an infected blood meal, of gametocytes, the precursor cells of the gametes. Relatively little is known about the molecular mechanisms involved in the developmental switch from asexual development to sexual differentiation or the maturation and survival of gametocytes. Here, we show that a gene coding for a novel putative transporter, NPT1, plays a crucial role in the development of Plasmodium berghei gametocytes. Parasites lacking NPT1 are severely compromised in the production of gametocytes and the rare gametocytes produced are unable to differentiate into fertile gametes. This is the earliest block in gametocytogenesis obtained by reverse genetics and the first to demonstrate the role of a protein with a putative transport function in sexual development. These results and the high degree of conservation of NPT1 in Plasmodium species suggest that this protein could be an attractive target for the development of novel drugs to block the spread of malaria.     

Demonstration of 5-Hydroxytryptamine in Blood Platelets by a Formaldehyde-Induced Fluorescence Technique

Blood smears from normal and reserpinized rabbits were dried in a desiccator and treated with gaseous formaldehyde. The normal blood smears showed an intense yellow fluorescence due to serotonin in platelets. Fluorescence was weak or absent in the smears of the reserpinized rabbits. A standard serotonin assay of the bloods confirmed a normal 5-hydroxytryptamine content in untreated rabbits while reserpinized rabbits had a low or zero 5-hydroxytryptamine level. This is a simple method for detecting the presence or absence of 5-hydroxytryptamine in the blood.     

鸡沙氏住白虫的生活史研究

本文报告了鸡沙氏住白虫生活史及其全程发育。经人工实验和流行区调查证明,后宽绳蚋是该虫的自然传播媒介。孢子增殖在后宽绳蚋体内、经3—4天完成发育。无性裂殖体增殖在鸡的肝、肾、心、肺、脾、脑和肠等内脏器官组织细胞内进行,经4—9天完成发育,该裂殖体有二型,即肝裂殖体和巨噬细胞裂殖体。感染9—13天后,血细胞内的配子体发育成熟。感染后第15—25天末稍血液中出现虫体的高峰期。